title: Interpreting dual ELISA and qPCR data for bacterial kidney disease of salmonids.
abstract: Although there are a variety of methods available for the detection of Renibacterium salmoninarum, the causative agent of bacterial kidney disease in salmon and trout, the enzyme-linked immunosorbent assay (ELISA) is probably the most widely used method. However, ELISA measures bacterial antigen, which does not necessarily reflect the number of cells present. We hypothesized that dual analysis of kidney tissue by ELISA and a quantitative real-time polymerase chain reaction assay (qPCR) would provide complementary information about antigen level and the number of bacterial genomes. We found that DNA extracted from the insoluble fraction of the ELISA tissue preparation produced the same qPCR result as DNA extracted directly from frozen tissue, permitting true dual analysis of the same tissue sample. We examined kidney tissue in this manner from individual free-ranging juvenile Chinook salmon and antibiotic-treated captive subadult Chinook salmon and observed 3 different patterns of results. Among the majority of fish, there was a strong correlation between the ELISA value and the qPCR value. However, subsets of fish exhibited either low ELISA values with elevated qPCR values or higher ELISA values with very low qPCR values. These observations suggest a conceptual model that allows inferences about the state of infection of individual fish based on dual ELISA/qPCR results. Although this model requires further assessment through experimental infections and treatments, it may have utility in broodstock selection programs that currently apply egg-culling practices based on ELISA alone.
authors.affiliation: Aquatic Farms, Ltd., 49-139 Kamehameha Highway, Kaneohe, Hawaii 96744, USA.
authors.forename: Shelly L
authors.initials: SL
authors.lastname: Nance
authors.forename: Michael
authors.initials: M
authors.lastname: Riederer
authors.forename: Tyler
authors.initials: T
authors.lastname: Zubkowski
authors.forename: Marc
authors.initials: M
authors.lastname: Trudel
authors.forename: Linda D
authors.initials: LD
authors.lastname: Rhodes
mesh_terms: D000193:Actinomycetales Infections / Q000382:microbiology / Q000662:veterinary*; D000818:Animals; D004269:DNA, Bacterial / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D005393:Fish Diseases / Q000382:microbiology*; D008835:Micrococcaceae / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012474:Salmon
chemical_list: D004269:DNA, Bacterial